Liu Kunmei, Wan Panpan, Huang Yue, Wang Bin, Wang Xuequan, Zhang Rui, Guo Le
Ningxia Key Laboratory of Cerebrocranial Disease, Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region 750004, P.R. China.
Department of Microbiology and Biochemical Pharmacy, School of Pharmacy, Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region 750004, P.R. China.
Exp Ther Med. 2022 Sep 13;24(5):670. doi: 10.3892/etm.2022.11607. eCollection 2022 Nov.
Astrocyte elevated gene-1 (AEG-1) is a key regulatory factor of progression in multiple types of tumor and neurodegenerative disease development. AEG-1 is associated with glutamate excitotoxicity due to its reported function of repressing excitatory amino acid transporter 2 expression in astrocytes. Although the function of AEG-1 has been demonstrated in neurological disorders, such as Alzheimer's disease and amyotrophic lateral sclerosis, the underlying mechanism of neuronal AEG-1 function remains unclear. The aim of the present study was to clarify the function and related mechanism of AEG-1 in neurons. A stable AEG-1-deficient HT22 neuronal cell line was constructed using CRISPR/Cas9 gene-editing technology. Reverse transcription-quantitative PCR and western blotting were carried out to analyze the knockdown efficiency of AEG-1-deficient HT22 cell line. RNA Sanger sequencing analysis was performed in AEG-1-deficient HT22 cells and wild-type HT22 cells without knockout (KO). Results from RNA sequencing revealed that AEG-1 modulated neuronal morphology and development by regulating the expression of numerous genes, such as ubiquitin C, C-X-C motif chemokine ligand 1, MMP9, Notch1, neuropilin 1 and ephrin type-A receptor 4. In addition, AEG-1 deficiency impacted several signaling pathways by mediating cell survival differentiation, apoptosis, and migration; this included the TNF-α pathway, the NF-κB pathway, the MAPK signaling pathway, the Notch signaling pathway and Axon guidance. Downregulation in cellular ion homeostasis, including ion channel function and neurotransmitter release, were observed after knocking out AEG-1 expression. Collectively, the present study provides insights into AEG-1-dependent gene regulation and signaling pathway transduction in neurons. The results of the present study may be applied for improving the understanding of AEG-1-associated central nervous system diseases.
星形胶质细胞上调基因1（AEG-1）是多种肿瘤进展和神经退行性疾病发展的关键调节因子。由于其在星形胶质细胞中抑制兴奋性氨基酸转运体2表达的功能，AEG-1与谷氨酸兴奋性毒性有关。尽管AEG-1的功能已在神经疾病如阿尔茨海默病和肌萎缩侧索硬化中得到证实，但其在神经元中的潜在功能机制仍不清楚。本研究的目的是阐明AEG-1在神经元中的功能及其相关机制。利用CRISPR/Cas9基因编辑技术构建了稳定的AEG-1基因敲除HT22神经元细胞系。通过逆转录定量PCR和蛋白质印迹法分析AEG-1基因敲除HT22细胞系的敲除效率。对AEG-1基因敲除的HT22细胞和未敲除的野生型HT22细胞进行RNA Sanger测序分析。RNA测序结果显示，AEG-1通过调节众多基因的表达来调控神经元的形态和发育，这些基因包括泛素C、C-X-C基序趋化因子配体1、基质金属蛋白酶9、Notch1、神经纤毛蛋白1和Ephrin A型受体4。此外，AEG-1基因敲除通过介导细胞存活分化、凋亡和迁移影响多个信号通路；这包括肿瘤坏死因子-α通路、核因子-κB通路、丝裂原活化蛋白激酶信号通路、Notch信号通路和轴突导向。敲除AEG-1表达后，观察到细胞离子稳态下调，包括离子通道功能和神经递质释放。总的来说，本研究为神经元中AEG-1依赖的基因调控和信号通路转导提供了见解。本研究结果可能有助于增进对AEG-1相关中枢神经系统疾病的理解。
